Skip to main content

Purification and properties of the GABA/benzodiazepine/barbiturate receptor protein from rat brain

  • Chapter
  • 106 Accesses

Abstract

The major postsynaptic receptor for the inhibitory neurotransmitter γ-aminobutyric acid (GABA) is a complex protein containing a chloride channel and modulatory sites for two classes of drugs, the benzodiazepines (BZ) and anti-convulsants/barbiturates (Olsen, 1982). These GABA receptors are also defined by sensitivity to the agonist muscimol and the antagonist bicuculline (Enna, 1983). Barbiturates and related depressants like pyrazolopyridines that enhance GABA-activated chloride permeability at the tissue and cellular level (Nicoll and colleagues, this volume) also are able to enhance benzodiazepine and GABA receptor binding at the molecular level (Leeb-Lundberg et al., 1980, 1981; Olsen & Snowman, 1982; Wong et al., 1984b). These in vitro barbiturate interactions are dependent on the presence of physiological levels of chloride or other anions able to permeate GABA ion channels, and are blocked by picrotoxin and related chloride channel antagonists, as well as by bicuculline, a GABA receptor antagonist.

This is a preview of subscription content, log in via an institution.

Buying options

Chapter
USD   29.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   84.99
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  • ASANO, T. & OGASAWARA, N. (1981). Chloridedependent stimulation of GABA and benzodiazepine receptor binding by pentobarbital. Brain Res., 225, 212–216.

    Article  PubMed  CAS  Google Scholar 

  • ASANO, T., YAMADA, Y. & OGASAWARA, N. (1983). Characterization of the solubilized GABA and benzodiazepine receptor from various regions of bovine brain. J. Neurochem., 40, 209–214.

    Article  PubMed  CAS  Google Scholar 

  • BARKER, J.C., GRATZ, E., OWEN, D.G. & STUDY, R.E. (1984). Pharmacological effects of clinically important drugs on the excitability of cultured mouse spinal neurons. In Actions and Interactions of GABA and Benzodiazepines. Bowery, N.G. (ed.) pp.203–216, New York: Raven Press.

    Google Scholar 

  • BRADFORD, M.M. (1976). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem., 72, 248–252.

    Article  PubMed  CAS  Google Scholar 

  • ENNA, S.J. (ed.) (1983). The GABA Receptors, Clifton, New Jersey: Humana Press.

    Google Scholar 

  • FROEHNER, S.C., REINESS, C.T. & HALL, Z.W. (1977). Subunit structure of the acetylcholine receptor from denervated rat skeletal muscle. J. biol. Chem., 252, 8589–8594.

    Google Scholar 

  • GAVISH, M. & SNYDER, S.H. (1981). λ-Aminobutyric acid and benzodiazepine receptors: copurification and characterization. Proc. natn. Acad. Sci. U.S.A., 78, 1939–1942.

    Google Scholar 

  • GREENLEE, D.V. & OLSEN, R.W. (1979). Solubilization of GABA receptor protein from mammalian brain. Biochem. biophys. Res. Commun., 88, 380–387.

    Google Scholar 

  • HARRISON, N.L. & SIMMONDS, M.A. (1983). Two distinct interactions of barbiturates and chlormethiazole with the GABA receptor complex in rat cuneate nucleus in vitro. Br. J. Pharmac., 80, 387–394.

    Google Scholar 

  • HUANG, L.Y. & BARKER, J.L. (1980). Pentobarbital: stereospecific actions of the (+) and (−) isomers

    Google Scholar 

  • revealed on cultured mammalian neurons. Science, 207, 195–197.

    Google Scholar 

  • KING, R.G. & OLSEN, R.W. (1984). Solubilization of convulsant/barbiturate binding activity on the GABA/benzodiazepine receptor complex. Biochem. biophys. Res. Commun., 119, 530–536.

    Google Scholar 

  • KNABE, V.J., RUMMEL, W., BUCH, H.P. & FRANZ, N. (1978). Optisch aktive Barbiturate. Synthese, Konfiguration and pharmacologische wirkung. Arzneimittel-Forschung, 28, 1048–1056.

    Google Scholar 

  • LEEB-LUNDBERG, F. & OLSEN, R.W. (1982). Interactions of barbiturates of various pharmacological categories with benzodiazepine receptors. Mol. Pharmac., 21, 320–328.

    Google Scholar 

  • LEEB-LUNDBERG, F., SNOWMAN, A. & OLSEN, R.W. (1980). Barbiturate receptors are coupled to benzodiazepine receptors. Proc. natn. Acad. Sci. U.S.A., 77, 7468–7472.

    Google Scholar 

  • LEEB-LUNDBERG, F., SNOWMAN, A. & OLSEN, R.W. (1981). Perturbation of benzodiazepine receptor binding by pyrazolopyridines involves picrotoxinin/barbiturate receptor sites. J. Neurosci., 1, 471–477.

    Google Scholar 

  • MACDONALD, R.L. & BARKER, J.L. (1978). Different actions of anticonvulsant and anesthetic barbiturates revealed by use of cultured mammalian spinal cord neurons: a common mode of anticonvulsant action. Brain Res., 167, 323–326.

    Google Scholar 

  • MOHLER, H. & OKADA, T. (1977). Benzodiazepine receptors: demonstration in the central nervous system. Science, 198, 849–851.

    Google Scholar 

  • MÖHLER, H., BATTERSBY, M.K. & RICHARDS, J.C. (1980). Benzodiazepine receptor protein identified and visualized in brain tissue by a photoaffinity label. Proc. natn. Acad. Sci. U.S.A., 77, 1666–1670.

    Google Scholar 

  • NICOLL, R.A., ECCLES, J.C., OSHIMA, T.C. & RUBIA, F. (1975). Prolongation of hippocampal inhibitory postsynaptic potentials by barbiturates. Nature, 258, 625–627.

    Google Scholar 

  • OLSEN, R.W. (1982). Drug interactions at the GABA receptor ionophore complex. A. Rev. Pharmac. Toxicol., 22, 245–277.

    Article  CAS  Google Scholar 

  • OLSEN, R. W., BERGMAN, M.O., VAN NESS, P.C., LUMMIS, S.C., WATKINS, A.E., NAPIAS, C. & GREENLEE, D.V. (1981). γ-Aminobutyric acid receptor binding in mammalian brain: heterogeneity of binding sites. Mol. Pharmac., 19, 217–227.

    Google Scholar 

  • OLSEN, R.W. & SNOWMAN, A.M. (1982). Chloride-dependent enhancement by barbiturates of GABA receptor binding. J. Neurosci., 2, 1812–1823.

    Google Scholar 

  • OLSEN, R.W. & SNOWMAN, A.M. (1983). [3H]Bicuculline methochloride binding to low affinity GABA receptor sites. J. Neurochem., 41. 1563–1663.

    Google Scholar 

  • OLSEN, R.W., FISCHER, J.B. & DUNWIDDIE, T.V. (1984a). Barbiturate enhancement of λ-aminobutyric acid receptor binding and function as a mechanism of anesthesia. In Molecular Mechanisms of Anaesthesia. Roth, S. & Miller, K. (eds) (in press).

    Google Scholar 

  • OLSEN, R.W., SNOWHILL, E.W. & WAMSLEY, J.K. (1984b). Autoradiographic localization of low affinity GABA receptors with [3H]bicuculline methochloride. Eur. J. Pharmac., 99, 247–248.

    Google Scholar 

  • OLSEN, R.W., WONG, E.H.F., STAUBER, G.B., MURAKAMI, D., KING, R.G. & FISCHER, J.B. (1984c). Biochemical Properties of the GABA/Barbiturate/Benzodiazepine Receptor-Chloride Ion Channel Complex. In Neurotransmitter Receptors: Mechanisms of Action and Regulation. Kito, S., Segawa, T., Kuriyarna, K., Yamamura, H.I. & Olsen, R.W. (eds) pp.205–219, New York: Plenum Press.

    Google Scholar 

  • SIGEL, E. & BARNARD, E.A. (1984). A λ-aminobutyric acid/benzodiazepine receptor complex from bovine cerebral cortex. Improved purification with preservation of regulatory sites and their interactions. J. biol. Chem., 259, 7219–7223.

    Google Scholar 

  • SIMMONDS, M.A. (1981). Distinction between the effects of barbiturates, benzodiazepines and phenytoin on responses to γ-aminobutyric acid receptor activation and antagonism by bicuculline and picrotoxinin. Br. J. Pharmac., 73, 739–747.

    Article  CAS  Google Scholar 

  • SQUIRES, R.F. & BRAESTRUP, C. (1977). Benzodiazepine receptors in rat brain. Nature, 266, 732–734.

    Google Scholar 

  • SQUIRES, R.F., CASIDA, J.E., RICHARDSON, M. & SAEDERUP, E. (1983). [35S]t-Butyl-bicyclophosphorothionate binds with high affinity to brain-specific sites coupled to γ-aminobutyric acid-A and ion recognition sites. Mol. Pharmac., 23, 326–336.

    Google Scholar 

  • STEPHENSON, F.A. & OLSEN, R.W. (1982). Solubilization by CHAPS detergent of barbiturate-enhanced benzodiazepine-GABA receptor complex. J. Neurochem., 39, 1579–1586.

    Google Scholar 

  • STEPHENSON, F.A., WATKINS, A.E. & OLSEN, R.W. (1982). Physicochemical characterization of detergent-solubilized γ-aminobutyric acid and benzodiazepine receptor protein from bovine brain. Eur. J. Biochem., 123, 291–298.

    Google Scholar 

  • SUPAVILAI, P. & KAROBATH, M. (1979). Stimulation of benzodiazepine receptor binding by SQ 20009 is chloride-dependent and picrotoxin-sensitive. Eur. J. Pharmac., 60, 111–113.

    Google Scholar 

  • SUPAVILAI, P., MANNONEN, A. & KAROBATH, M. (1982). Modulation of GABA binding sites by CNS depressants and CNS convulsants. Neurochem. Int., 4, 259–268.

    Google Scholar 

  • TALLMAN, J.F., PAUL, S.M., SKOLNICK, P. & GALLAGER, D.W. (1980). Receptors for the age of anxiety: pharmacology of the benzodiazepines. Science, 207, 274–281.

    Google Scholar 

  • TICKU, M.K., BAN, M. & OLSEN, R.W. (1978). Binding of [3H]α-dihydropicrotoxinin, a γ-aminobutyric acid synaptic antagonist, to rat brain membranes. Mol. Pharmac., 14, 391–402.

    Google Scholar 

  • TICKU, M.K. & MAKSAY, G. (1983). Convulsant/depressant site of action at the allosteric benzodiazepine-GABA receptor-ionophore complex. Life Sci., 33, 2263–2375.

    Google Scholar 

  • TICKU, M.K. & OLSEN, R.W. (1978). Interaction ot barbiturates with dihydropicrotoxin in binding sites related to the GABA receptor-ionophore system. Life Sci., 22, 1643–1651.

    Google Scholar 

  • WHITTLE, S.R. & TURNER, A.J. (1982). Differential effects of sedative and anticonvulsant barbiturates on specific [3H]GABA binding to membrane preparations from rat brain cortex. Biochem. Pharmac., 31, 2891–2895.

    Google Scholar 

  • WILLOW, M. & JOHNSTON, G.A.R. (1980). Enhancement of GABA binding by pentobarbitone. Neurosci. Lett., 18, 323–327.

    Article  PubMed  CAS  Google Scholar 

  • WONG, E.H.F., LEEB-LUNDBERG, L.M.F., TEICHBERG, V.I. & OLSEN, R.W. (1984a). γ-Aminobutyric acid activation of 36Cl flux in rat hippocampal slices and its potentiation by barbiturates. Brain Res., 303, 267–275.

    Article  PubMed  CAS  Google Scholar 

  • WONG, E.H.F., SNOWMAN, A.M., LEEB-LUNDBERG, L.M.F. & OLSEN, R.W. (1984b). Barbiturates inhibit GABA antagonist and benzodiazepine inverse agonist binding. Eur. J. Phanmac. (in press).

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

William Paton James Mitchell Paul Turner Cheryl Padgham Eileen Ashcroft

Copyright information

© 1984 Macmillan Publishers Limited

About this chapter

Cite this chapter

Olsen, R.W., Fischer, J.B., Stauber, G.B., King, R.G., Yang-Ransom, J. (1984). Purification and properties of the GABA/benzodiazepine/barbiturate receptor protein from rat brain. In: Paton, W., Mitchell, J., Turner, P., Padgham, C., Ashcroft, E. (eds) IUPHAR 9th International Congress of Pharmacology London 1984. Palgrave Macmillan, London. https://doi.org/10.1007/978-1-349-17615-1_25

Download citation

Publish with us

Policies and ethics